G Protein-coupled Receptors: Molecular Pharmacology From Academic Concept to Pharmaceutical Research

284 Pages·2008·10.55 MB·English

by  Georges Vauquelin

G Protein-coupled Receptors
G Protein-Coupled Receptors: Molecular Pharmacology From Academic Concept to Pharmaceutical Research
Georges Vauquelin and Bengt von Mentzer © 2007 John Wiley & Sons, Ltd. ISBN: 978-0-470-51647-8
G Protein-coupled Receptors
Molecular Pharmacology
From Academic Concept to
Pharmaceutical Research
Georges Vauquelin
Institute for Molecular Biology and Biotechnology,
Free University Brussels, Belgium
Bengt von Mentzer
GI Molecular Pharmacology, AstraZeneca R&D Mölndal, Sweden
Copyright © 2007 John Wiley & Sons Ltd, The Atrium, Southern Gate, Chichester,
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Library of Congress Cataloguing-in-Publication Data
Vauquelin, Georges.
G protein-coupled receptors : molecular pharmacology/
Georges Vauquelin and Bengt von Mentzer.
p. ; cm.
Includes bibliographical references and index.
ISBN 978-0-470-51647-8
1. G proteins–Receptors. 2. Molecular pharmacology. I. Mentzer, Bengt
von. II. Title.
[DNLM: 1. Receptors, G Protein-Coupled–physiology. 2. Drug Design.
3. GTP-Binding Proteins–physiology. 4. Signal
Transduction–physiology. QU 55.7 V378g 2007]
QP552.G16V38 2007
615(cid:2).1–dc22 2007024241
British Library Cataloguing in Publication Data
A catalogue record for this book is available from the British Library
ISBN: 978-0-470-51647-8
Typeset in 10.5/12.5 pt Times by Thomson Digital
Printed and bound in Trento, Italy by Printer Trento S.r.l.
This book is printed on acid-free paper responsibly manufactured from sustainable forestry in which at least
two trees are planted for each one used for paper production.
GEORGES VAUQUELIN BENGT VON MENTZER
Institute for Molecular Biology and Biotechnology, GI Molecular Pharmacology, AstraZeneca
Free University Brussels, Belgium. R&D Mölndal, Sweden.
Prof. Vauquelin’s initial research dealt with the purifi cation of β-adrenergic receptors
(PhD, 1978) and the discrimination between agonist- and antagonist- β-adrenergic
receptor interaction. Part of this work was performed in 1979-1980 at the Case West-
ern Reserve University, Cleveland, during a post-doctoral stay in the lab of Prof. M.
Maguire. Dr. Vauquelin’s subsequent work focused on in vitro techniques. It included
the identifi cation and characterization of GPCRs and mechanisms of receptor regula-
tion by physiological and patho-physiological conditions with a particular emphasis on
receptors in the human CNS. Some of his research also dealt with the interaction of
natural products from plants and animal venoms (conotoxins) with receptors and their
ligands and with the investigation of membrane proteins with potential receptor-like
activity. More recently, intact cell systems expressing either endogenous or transfected
receptors (for neuropeptide Y, angiotensin II) have been used to compare ligand-recep-
tor interactions by radioligand binding and functional measurements. Dr. Vauquelin is
(co)author of over 180 publications in international journals.
Dr. von Mentzer’s initial research dealt with mechanisms, classifi cation and locali-
zation of β-adrenergic receptors involved in colon motility (PhD, 1985). Dr. von
Mentzer’s postdoctoral studies were focused on the understanding of the functional
impact of drug-receptor interactions at the University of Leicester with Prof. Stefan
Nahorski. These studies comprised the functional coupling of muscarinic receptors
to phosphoinositol signalling pathways. During the ensuing 10 years, Dr. von Mentzer
moved to the fi eld of fatty acid metabolism, concentrating on the arachidonic acid
pathway. These studies led to a joint collaboration with Prof. Arthur Spector at the
University of Iowa and Prof. Jim Hamilton at Boston University. Dr. von Mentzer re-
turned back to receptor pharmacology and developed a particular interest in neuropep-
tide research. His present occupation deals with drug development at AstraZeneca.
He is an initiator of teamwork in several discovery/development projects related to
gastrointestinal neuropeptide GPCRs and provides scientifi c support to project and
group leaders at different stages in drug development.
Thinking must never submit itself,
neither to a dogma,
nor to a party,
nor to a passion,
nor to an interest,
nor to a preconceived idea,
nor to anything whatsoever,
except to the facts themselves,
because for it to submit to anything else would be the end of its existence.
Henri Poincaré (French mathematician and natural philosopher 1854–1912)
To Bengt Åblad and Enar Carlsson who introduced us into their world of knowledge,
enthusiasm and wisdom, and greatly stimulated our eagerness to undertake this
enterprise. Many thanks from both authors.
Contents
Preface xi
Foreword xii
1 Chemical messengers and the cell membrane 1
1.1 Endocrine signalling by hormones 1
1.2 The nervous system and synaptic signalling by neurotransmitters 8
Small molecule neurotransmitters 11
Neuropeptides 13
1.3 Paracrine signalling by local chemical messengers 15
1.4 Hydrophobicity: effect on release and transport of messengers 16
1.5 Membrane proteins and membrane receptors 23
1.6 Ligand–receptor interactions 25
2 Radioliogand binding studies 29
2.1 Technical aspects of radioligand binding 29
2.2 Saturation binding 33
2.3 Competition binding 38
2.4 Kinetic experiments 46
2.5 Regional distribution of receptors 50
3 Functional studies 53
3.1 Dose-response curves and associated problems 53
3.2 From receptor occupation to stimulus and response 56
From receptor occupation to stimulus 56
From stimulus to response: linear relationship 59
From stimulus to response: non-linear relationship 61
3.3 Receptor classifi cation and antagonist affi nity 65
3.4 Pharmacological models 69
4 G protein-coupled receptors 77
4.1 From receptor to response: introduction to GPCRs 77
4.2 GPCR structure 86
4.3 Ligand interactions with family A, B and C receptors 97
Ligand interaction with family A receptors 98
CONTENTS
Ligand interaction with family B receptors 102
Ligand interaction with family C receptors 104
4.4 Receptor activation 106
4.5 Activated GPCRs: interaction with G proteins 113
Techniques to study G protein-coupling preference 119
Divergence of intracellular signalling 123
4.6 Activated GPCRs: phosphorylation and internalization 125
Receptor phosphorylation 125
β-Arrestin binding mediated GPCR endocytosis 129
Mechanisms to terminate signalling 134
4.7 β-Arrestin-binding and MAP kinase activation 137
4.8 GPCR dimerization and association with other proteins 142
Introductory comments 142
GPCR dimerization 143
GPCR interaction with other membrane receptors 151
GPCR interaction with other membrane proteins 152
GPCR interactions with cytoplasmic proteins 156
4.9 Early models for GPCR activation 159
4.10 Restricted GPCR mobility and G protein coupling 163
Membrane compartimentalization 163
Restricted GPCR–G protein coupling: effector activity 173
4.11 Spontaneous receptor–G protein coupling 175
Models 175
Inverse agonism 179
4.12 Interaction of two G proteins with one activated receptor state 183
Fusion proteins between GPCRs and G proteins 185
4.13 Multiple receptor conformations 191
‘Agonist traffi cking’: what do models predict? 196
Experimental ‘evidence’ for agonist traffi cking: potential pitfalls 198
Multistate receptors: ligand-mediated sequential changes in
receptor conformation 201
Multiple receptor states related to truncation, covalent modifi cation
and mutation 204
4.14 Multistate receptors and multiple ligand binding sites 207
The general allosteric ternary complex model 210
Exogenous and endogenous allosteric modulators 212
allosteric phenomena at GPCR: detection by radioligand binding 214
Detection of allosteric phenomena at GPCRs by functional assays 219
Usefulness of allosteric modulators 222
4.15 ‘Competitive’, ‘non-competitive’ and ‘insurmountable’ antagonism 222
Co-incubation, no receptor reserve 222
Antagonist pre-incubation, no receptor reserve 224
4.16 Naturally occurring mutations of GPCRs 226
5 Concluding remarks 231
References 235
Index 244
Preface
This book provides, in the fi rst place, useful scientifi c background to those involved
in pre-clinical research and to those who need to function within multi-disciplinary
teams in the pharmaceutical industry (from medical chemistry and molecular biology
via pharmacology to drug marketing). Today’s education of researchers in the
pharmaceutical industry needs to be broad and insight into pharmacological issues
needs to be warranted. In this respect, the focus on G protein-coupled receptors
(GPCRs) is rationalised by the fact that they constitute major cellular targets for
pharmaceutical drugs.
The aim is also to provide students and investigators with a basic interest in life
sciences an insight into the fast evolving GPCR fi eld. More generally speaking, to
create knowledge in the domain of molecular pharmacology. In this respect, the book
will also provide necessary background for the elaboration and reading of research
papers on GPCRs and even information brochures about pharmaceutical drugs (which
nowadays often describe their molecular mechanism of action).
To these ends:
1. much attention is devoted to the analysis and interpretation of experimental data
(radioligand binding and functional assays)
2. the now widespread use of recombinant cell lines, receptor mutants and related
artifi ces in drug research is critically evaluated
3. special attention is also devoted to trendy but often poorly understood concepts
such as insurmountable antagonism, inverse agonism and allosteric interactions.
We are especially indebted to Ulla Schefström, Lena Ruehl and Ann-Marie Ashton
for their excellent administrative and technical support.
Georges Vauquelin and Bengt von Mentzer
July 2007
Foreword
The greatest need for new textbooks occurs in areas that are rapidly changing. Such
textbooks have to be written by scientists actually involved in the research which is
advancing the fi eld. It is most often very diffi cult to persuade experts in these fi elds
to set time aside to spread their knowledge and to produce concise but instructive
information. Georges Vauquelin and Bengt von Mentzer have a recognized expertise
on G protein-coupled receptors. Their book on this matter is, therefore, unique. It is an
updated work that covers the major aspects of the present knowledge concerning this
very interesting class of receptors.
The science of GPCRs is one of the most attractive areas of modern biological
research. As it is growing very rapidly, this book will be very useful for many workers
in biological as well as clinical science dealing with these receptors. It should even
help to stimulate new lines of research.
GPCRs are widely considered as interesting targets for drugs. In fact, some of them
are already used as targets for very popular drugs such as beta-blockers, muscarinic
agonists and antagonists, many psychotropic drugs and others. Very intense research
on GPCRs is being conducted in academic as well as in pharmaceutical industry
laboratories dealing with drug design and pharmaceutical innovation.
As a pharmacologist involved in experimental and clinical research, I consider this
book to be essential reading. It should serve the needs of scientists of course, but also of
graduate and postgraduate students in pharmacology, biochemistry and cell biology.
Pascal Bousquet
Professor of Pharmacology Medical School
Louis Pasteur University
Strasbourg
France